PROJECT SUMMARY/ABSTRACT Nearly one-third of the world population is infected with Mycobacterium tuberculosis. This reservoir contributes towards an increasing incidence of tuberculosis, with about 11 million new cases and 1.8 million deaths every year. The emergence of multi-drug resistant strains combined with low diagnostic and therapeutic coverage in many developing countries, has worsened the disease burden caused by TB. The profound success of Mycobacterium tuberculosis in causing disease depends on its ability to successfully utilize the host?s cellular machineries including the lipid biosynthesis pathway to subvert the immune system. Our understanding of these processes especially that of the host lipid biology in the context of Mycobacterium tuberculosis infection is limited. The aim of the proposed work is to study the role of host?s lipid biosynthesis in the entry, replication and killing of Mycobacterium tuberculosis. For this purpose, individual knockout macrophage cell lines that lack genes involved in the biosynthesis of lipids will be generated using CRISPR/Cas9-technology. Understanding these pathways or processes essential for the life cycle of Mycobacterium tuberculosis is crucial, as they represent potential targets for new therapeutics.